OBJECTIVE: The objective of this study was to evaluate the in vitro antifungal activity of propolis extract against yeasts Candida albicans and Candida non-albicans isolated from vaginal exudates, in comparison with nystatin. DESIGN: Ninety-seven (97) vaginal yeasts strains were evaluated. These strains were obtained from different clinical conditions, isolated and stored at the Sector of Medical Mycology of the State University of Maringá (Paraná, Brazil). The assays of susceptibility to nystatin and propolis extracts (PE) were conducted through microdilution in broth (National Committee for Clinical Laboratory Standards-NCCLS, M-27A Document of 1997). RESULTS: All the yeasts tested were inhibited by low concentrations of PE (maximum of 393.19 mug/mL of the total flavonoid content), including an isolate resistant to nystatin, regardless of the clinical conditions of the women and the species of yeast isolated. CONCLUSIONS: The PE showed an outstanding performance against the tested vaginal yeast strains, and could be included among the novel therapeutic options for the treatment of vulvovaginal candidiasis.